I am using wortmannin as an inhibitor of autophagy. When I check autophagy inhibition by WB of LC3-II, in some cell lines wortmannin treated cells have more LC3-II than untreated, contrarily to what is expected. Any idea about why this is happening?
Does chloroquine or other autophagy inhibitor act in the same way in those cell lines? You also should do the kinetics of LC3B accumulation. Sometimes the single timepoint only gives you a static picture that is not representing the whole process. Hope it helps.
Which concantartion have you used? Is it specific for Wortmanin, or you observde a non-specfic binding and non-specfci effect because of wrong concnatration? Good luck!
Hi, Gemma, in addition to previous post, please, consider that each compounds have specific "pharmacological" window which dependent from the concentration and cell types/conditions. If you go out from this specific windows, you can get a couple of not-specific effects whcih does not related directly with the compound specific action. Kinetics, as Vadim mention previously is also very important!
I agree with Taras. You have to take into consideration of the context. Are you studying basal autophagy or starvation induced autophagy? Dose of the drug is also important since too much will induce some off target response.
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